Pharmaceutical compositions and methods of treating dry eye disorders

ABSTRACT

Ophthalmic compositions comprising Inhibitors of Janus kinase-3 (“Jak3”) are useful for treating dry eye disorders and other disorders requiring the wetting of the eye.

This application is a divisional of U.S. application Ser. No.12/688,604, which is a National Stage Application under 35 U.S.C. §371of PCT/IB2008/001819, filed on Jul. 4, 2008, which claims the benefit ofU.S. Provisional Application No. 60/949,216, filed on Jul. 11, 2007 andU.S. Provisional Application No. 61/060,032, filed on Jun. 9, 2008, theentire contents of which are incorporated herein by reference.

BACKGROUND OF THE INVENTION

Dry eye, also known generically as keratoconjunctivitis sicca, is acommon ophthalmological disorder affecting millions of Americans eachyear. Dry eye is a multifactorial disease of the tears and ocularsurface that results in symptoms of discomfort, visual disturbance, andtear film instability with potential damage to the ocular surface. It isaccompanied by increased osmolarity of the tear film and inflammation ofthe ocular surface.

Dry eye may afflict an individual with varying severity. Dry eye isparticularly widespread among post-menopausal women due to hormonalchanges following the cessation of fertility. In mild cases, a patientmay experience burning, a feeling of dryness, and persistent irritationsuch as is often caused by small bodies lodging between the eye lid andthe eye surface. In severe cases, vision may be substantially impaired.Other diseases, such as Sjogren's disease and cicatricial pemphigoidmanifest dry eye complications.

The etiopathogenic classification of dry eye was first developed in 1995by the National Eye Institute/Industry Dry Eye Workshop and was furtherupdated in the January 2007 definition. The Ocular Surface, vol. 5, no.2, 75-92 (2007). The major classes of dry eye include aqueoustear-deficient dry eye and evaporative dry eye. Aqueous tear-deficientdry eye comprises two major subclasses, Sjogren syndrome dry eye andnon-Sjogren syndrome dry eye (primary and secondary lacrimal glanddeficiencies, obstruction of the lacrimal gland ducts, reflexhyposecretion, reflex motor block). Evaporative dry eye, resulting fromexcessive water loss from the exposed ocular surface in the presence ofnormal lacrimal secretory function, comprises the subclasses ofintrinsic causes (meibomian gland dysfunction, lid aperture andlid/globe disorders, low blink rate) and extrinsic causes (ocularsurface disorders and disease, contact lens wear, allergicconjunctivitis such as vernal keratoconjunctivitis).

Although it appears that dry eye may result from a number of unrelatedpathogenic causes, all presentations of the complication share a commoneffect, that is the breakdown of the pre-ocular tear film, which resultsin dehydration of the exposed outer surface and many of the symptomsoutlined above (Lemp, Report of the national Eye Institute/Industryworkshop on Clinical Trials in Dry Eyes, the CLAO Journal, vol. 21, no.4, pp 221-231 (1995).

Practitioners have taken several approaches to the treatment of dry eye.One common approach has been to supplement and stabilize the ocular tearfilm using so-called artificial tears instilled throughout the day.Other approaches include the use of ocular inserts that provide a tearsubstitute or stimulation of endogenous tear production.

Examples of the tear substitution approach include the use of buffered,isotonic saline solutions, aqueous solutions containing water solublepolymers that render the solutions more viscous and thus less easilyshed by the eye. Tear reconstitution is also attempted by providing oneor more components of the tear film such as phospholipids and oils.Phospholipid compositions have been shown to be useful in treating dryeye; see, e.g. McCulley and Shine, Tear film structure and dry eye,Contactologio, vol. 20(4), pp 145-9 (1998). Phosphilipid drug deliverysystems involving phospholipids, propellants and an active substance arealso known, see U.S. Pat. No. 5,174,988.

Another approach involves the provision of lubricating substances inlieu of artificial tears. For example, U.S. Pat. No. 4,818,537 (Guo)discloses the use of a lubricating liposome-based composition, and U.S.Pat. No. 5,800,807 (Hu et al.) discloses compositions containingglycerin and propylene glycol for treating dry eye.

Although these approaches have met with some success, problems in thetreatment of dry eye nevertheless remain. The use of tear substitutes,while temporarily effective, generally requires repeated applicationover the course of a patient's waking hours. It is not uncommon for apatient to have to apply artificial tear solution ten to twenty timesover the course of the day. Such an undertaking is not only cumbersomeand time consuming, but is also potentially very expensive. Transientsymptoms of dry eye associated with refractive surgery have beenreported to last in some cases from six weeks to six months or morefollowing surgery.

Aside from efforts directed primarily to the alleviation of symptomsassociated with dry eye, methods and compositions directed to treatmentof the dry eye condition have also been pursued. For example, U.S. Pat.No. 5,041,434 (Lubkin) discloses the use of sex steroids such asconjugated estrogens to treat dry eye conditions in post-menopausalwomen; U.S. Pat. No. 5,290,572 (MacKeen) discloses the use of finelydivided calcium ion compositions to stimulate pre-ocular tear filmproduction; and U.S. Pat. No. 4,966,773 (Gressel et al.) discloses theuse of microfine particles of one or more retinoids for ocular tissuenormalization.

Some literature reports suggest that patients suffering from dry eyesyndrome disproportionately exhibit the hallmarks of excessiveinflammation in relevant ocular tissues, such as the lacrimal andmeibomian glands. The use of various compounds to treat dry eyepatients, such as steroids e.g. U.S. Pat. No. 5,968,912; Marsh, et al.,Topical nonpreserved methylprednisolone therapty forkeratoconjunctivitis sicca in Sjogren syndrome, Ophthalmology, 106(5):881-816 (1999); Plugfelder et al, U.S. Pat. No. 6,153,607]; cytokinerelease inhibitors (Yanni, J. M.; et al WO0003705 A1), cyclosporine A[Tauber, J. Adv. Exp. Med. Biol. 1998, 438, (Lacrimal Gland, Tear Fromand Dry Eye Syndromes 2), 969] and 15-HETE (Yanni et al., U.S. Pat. No.5,696,166) has been disclosed.

In addition to dry eye, Jak3 inhibitors may be useful in the treatmentof other inflammation-linked ocular disorders, including but not limitedto glaucoma, uveitis, diabetic retinopathy and age-related maculardegeneration. A CNIB-funded study found that patients having theinflammatory biomarker, anticardiolipin, were four times more likely toprogress in glaucoma. Jak3 inhibitors have previously been suggested forthe treatment of diabetes, although there appears no suggestion fordiabetic retinopathy in particular. Cetkovic-Cvrle, M. and Uckun, F. M.,Arch Immunol Ther Exp (Warsz), 52(2), 69-82 (2004). Rodrigues found thatthe isolation of immunoglobulins, complement proteins, cytokines andactivated microglia, in retinal pigment epithelium (RPE) cells anddrusen provided evidence for the role of inflammation in dry age-relatedmacular degeneration. Rodrigues E. B., Ophthalmologica, 221(3):143-52(2007).

SUMMARY OF THE INVENTION

The present invention is directed to a method for the treatment of dryeye which comprises administering to a mammal a composition comprising apharmaceutically acceptable carrier and a pharmaceutically effectiveamount of a Janus kinase-3 (“Jak3”) inhibitor. In one aspect, the Jak3inhibitors have the compound of formula I:

wherein

R¹ is a group of the formula

wherein y is 0, 1 or 2;

R⁴ is selected from the group consisting of hydrogen, (C₁-C₆)alkyl,(C₁-C₆)alkylsulfonyl, (C₂-C₆)alkenyl, (C₂-C₆)alkynyl wherein the alkyl,alkenyl and alkynyl groups are optionally substituted by deuterium,hydroxy, amino, trifluoromethyl, (C₁-C₄)alkoxy, (C₁-C₆)acyloxy,(C₁-C₆)alkylamino, ((C₁-C₆)alkyl)₂amino, cyano, nitro, (C₂-C₆)alkenyl,(C₂-C₆)alkynyl or (C₁-C₆)acylamino; or R⁴ is (C₃-C₁₀)cycloalkyl whereinthe cycloalkyl group is optionally substituted by deuterium, hydroxy,amino, trifluoromethyl, (C₁-C₆)acyloxy, (C₁-C₆)acylamino,(C₁-C₆)alkylamino, ((C₁-C₆)alkyl)₂amino, cyano, cyano(C₁-C₆)alkyl,trifluoromethyl(C₁-C₆)alkyl, nitro, nitro(C₁-C₆)alkyl or(C₁-C₆)acylamino;

R⁵ is (C₂-C₉)heterocycloalkyl wherein the heterocycloalkyl groups mustbe substituted by one to five carboxy, cyano, amino, deuterium, hydroxy,(C₁-C₆)alkyl, (C_(r) C₆)alkoxy, halo, (C₁-C₆)acyl, (C₁-C₆)alkylamino,amino(C₁-C₆)alkyl, (C₁-C₆)alkoxy-CO—NH, (C₁-C₆)alkylamino-CO—,(C₂-C₆)alkenyl, (C₂-C₆)alkynyl, (C₁-C₆)alkylamino, amino(C₁-C₆)alkyl,hydroxy(C₁-C₆)alkyl, (C₁-C₆)alkoxy(C₁-C₆)alkyl,(C₁-C₆)acyloxy(C₁-C₆)alkyl, nitro, cyano(C₁-C₆)alkyl, halo(C₁-C₆)alkyl,nitro(C₁-C₆)alkyl, trifluoromethyl, trifluoromethyl(C₁-C₆)alkyl,(C₁-C₆)acylamino, (C₁-C₆)acylamino(C₁-C₆)alkyl,(C₁-C₆)alkoxy(C₁-C₆)acylamino, amino(C₁-C₆)acyl,amino(C₁-C₆)acyl(C₁-C₆)alkyl, (C₁-C₆)alkylamino(C₁-C₆)acyl,((C₁-C₆)alkyl)₂amino(C₁-C₆)acyl, R¹⁵R¹⁶N—CO—O—, R¹⁵R¹⁶N—CO—(C₁-C₆)alkyl,(C₁-C₆)alkyl-S(O)_(m), R¹⁵R¹⁶NS(O)_(m), R¹⁵R¹⁶NS(O)_(m) (C₁-C₆)alkyl,R¹⁵S(O)_(m)R¹⁶N, R¹⁵S(O)_(m)R¹⁶N(C₁-C₆)alkyl wherein m is 0, 1 or 2 andR¹⁵ and R¹⁶ are each independently selected from hydrogen or(C₁-C₆)alkyl; or a group of the formula

wherein a is 0, 1, 2, 3 or 4;b, c, e, f and g are each independently 0 or 1;d is 0, 1, 2, or 3;X is S(O)_(n) wherein n is 0, 1 or 2; oxygen, carbonyl or —C(═N-cyano)-;Y is S(O)_(n) wherein n is 0, 1 or 2; or carbonyl; andZ is carbonyl, C(O)O—, C(O)NR— or S(O)_(n) wherein n is 0, 1 or 2;

R⁶, R⁷, R⁸, R⁹, R¹⁰ and R¹¹ are each independently selected from thegroup consisting of hydrogen or (C₁-C₆)alkyl optionally substituted bydeuterium, hydroxy, amino, trifluoromethyl, (C₁-C₆)acyloxy,(C₁-C₆)acylamino, (C₁-C₆)alkylamino, ((C₁-C₆)alkyl)₂amino, cyano,cyano(C₁-C₆)alkyl, trifluoromethyl(C₁-C₆)alkyl, nitro, nitro(C₁-C₆)alkylor (C₁-C₆)acylamino;

R¹² is carboxy, cyano, amino, oxo, deuterium, hydroxy, trifluoromethyl,(C₁-C₆)alkyl, trifluoromethyl(C₁-C₆)alkyl, (C₁-C₆)alkoxy, halo,(C₁-C₆)acyl, (C₁-C₆)alkylamino, ((C₁-C₆)alkyl)₂ amino,amino(C₁-C₆)alkyl, (C₁-C₆)alkoxy-CO—NH, (C₁-C₆)alkylamino-CO—,(C₂-C₆)alkenyl, (C₂-C₆)alkynyl, (C₁-C₆)alkylamino, hydroxy(C₁-C₆)alkyl,(C₁-C₆)alkoxy(C₁-C₆)alkyl, (C₁-C₆)acyloxy(C₁-C₆)alkyl, nitro,cyano(C₁-C₆)alkyl, halo(C₁-C₆)alkyl, nitro(C₁-C₆)alkyl, trifluoromethyl,trifluoromethyl(C₁-C₆)alkyl, (C₁-C₆)acylamino,(C₁-C₆)acylamino(C₁-C₆)alkyl, (C₁-C₆)alkoxy(C₁-C₆)acylamino,amino(C₁-C₆)acyl, amino(C₁-C₆)acyl(C₁-C₆)alkyl,(C₁-C₆)alkylamino(C₁-C₆)acyl, ((C₁-C₆)alkyl)₂amino(C₁-C₆)acyl,R¹⁵R¹⁶N—CO—O—, R¹⁵R¹⁶N—CO—(C₁-C₆)alkyl, R¹⁵C(O)NH, R¹⁵OC(O)NH,R¹⁵NHC(O)NH, (C₁-C₆)alkyl-S(O)_(m), (C₁-C₆)alkyl-S(O)_(m)—(C₁-C₆)alkyl,R¹⁵R¹⁶NS(O)_(m), R¹⁵R¹⁶NS(O)_(m) (C₁-C₆)alkyl, R¹⁵S(O)_(m)R¹⁶N,R¹⁵S(O)_(m)R¹⁶N(C₁-C₆)alkyl wherein m is 0, 1 or 2 and R¹⁵ and R¹⁶ areeach independently selected from hydrogen or (C₁-C₆)alkyl;

R² and R³ are each independently selected from the group consisting ofhydrogen, deuterium, amino, halo, hydroxy, nitro, carboxy,(C₂-C₆)alkenyl, (C₂-C₆)alkynyl, trifluoromethyl, trifluoromethoxy,(C₁-C₆)alkyl, (C₁-C₆)alkoxy, (C₃-C₁₀)cycloalkyl wherein the alkyl,alkoxy or cycloalkyl groups are optionally substituted by one to threegroups selected from halo, hydroxy, carboxy, amino(C₁-C₆)alkylthio,(C₁-C₆)alkylamino, ((C₁-C₆)alkyl)₂amino, (C₅-C₉)heteroaryl,(C₂-C₉)heterocycloalkyl, (C₃-C₉)cycloalkyl or (C₆-C₁₀)aryl; or R² and R³are each independently (C₃-C₁₀)cycloalkyl, (C₃-C₁₀)cycloalkoxy,(C₁-C₆)alkylamino, ((C₁-C₆)alkyl)₂amino, (C₆-C₁₀)arylamino,(C₁-C₆)alkylthio, (C₆-C₁₀)arylthio, (C₁-C₆)alkylsulfinyl,(C₆-C₁₀)arylsulfinyl, (C₁-C₆)alkylsulfonyl, (C₆-C₁₀)arylsulfonyl,(C₁-C₆)acyl, (C₁-C₆)alkoxy-CO—NH—, (C₁-C₆)alkyamino-CO—,(C₅-C₉)heteroaryl, (C₂-C₉)heterocycloalkyl or (C₆-C₁₀)aryl wherein theheteroaryl, heterocycloalkyl and aryl groups are optionally substitutedby one to three halo, (C₁-C₆)alkyl, (C₁-C₆)alkyl-CO—NH—,(C₁-C₆)alkoxy-CO—NH—, (C₁-C₆)alkyl-CO—NH—(C₁-C₆)alkyl,(C₁-C₆)alkoxy-CO—NH—(C₁-C₆)alkyl, (C₁-C₆)alkoxy-CO—NH—(C₁-C₆)alkoxy,carboxy, carboxy(C₁-C₆)alkyl, carboxy(C₁-C₆)alkoxy,benzyloxycarbonyl(C₁-C₆)alkoxy, (C₁-C₆)alkoxycarbonyl(C₁-C₆)alkoxy,(C₆-C₁₀)aryl, amino, amino(C₁-C₆)alkyl, (C₁-C₆)alkoxycarbonylamino,(C₆-C₁₀)aryl(C₁-C₆)alkoxycarbonylamino, (C₁-C₆)alkylamino,((C₁-C₆)alkyl)₂amino, (C₁-C₆)alkylamino(C₁-C₆)alkyl,((C₁-C₆)alkyl)₂amino(C₁-C₆)alkyl, hydroxy, (C₁-C₆)alkoxy, carboxy,carboxy(C₁-C₆)alkyl, (C₁-C₆)alkoxycarbonyl,(C₁-C₆)alkoxycarbonyl(C₁-C₆)alkyl, (C₁-C₆)alkoxy-CO—NH—,(C₁-C₆)alkyl-CO—NH—, cyano, (C₅-C₉)heterocycloalkyl, amino-CO—NH—,(C₁-C₆)alkylamino-CO—NH—, ((C₁-C₆)alkyl)₂amino-CO—NH—,(C₆-C₁₀)arylamino-CO—NH—, (C₅-C₉)heteroarylamino-CO—NH—,(C₁-C₆)alkylamino-CO—NH—(C₁-C₆)alkyl, ((C₁-C₆)alkyl)₂amino-CO—NH—(C₁-C₆)alkyl, (C₆-C₁₀)arylamino-CO—NH—(C₁-C₆)alkyl,(C₅-C₉)heteroarylamino-CO—NH—(C₁-C₆)alkyl, (C₁-C₆)alkylsulfonyl,(C₁-C₆)alkylsulfonylamino, (C₁-C₆)alkylsulfonylamino(C₁-C₆)alkyl,(C₆-C₁₀)arylsulfonyl, (C₆-C₁₀)arylsulfonylamino,(C₆-C₁₀)arylsulfonylamino(C₁-C₆)alkyl, (C₁-C₆)alkylsulfonylamino,(C₁-C₆)alkylsulfonylamino(C₁-C₆)alkyl, (C₅-C₉)heteroaryl or(C₂-C₉)heterocycloalkyl, or the pharmaceutically acceptable saltsthereof.

The present invention is also directed to a method for the treatment ofglaucoma, uveitis, diabetic retinopathy and age-related maculardegeneration comprising administering to a mammal a compositioncomprising a pharmaceutically effective amount of a Jak3 inhibitorhaving the compound of formula I.

Specific compounds of formula I include those wherein said compound isselected from the group consisting of:

-   Methyl-[4-methyl-1-(propane-1-sulfonyl)-piperidin-3-yl]-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amine;-   4-Methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidine-1-carboxylic    acid methyl ester;-   3,3,3-Trifluoro-1-{4-methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidin-1-yl}-propan-1-one;-   4-Methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidine-1-carboxylic    acid dimethylamide;-   ({4-Methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidine-1-carbonyl}-amino)-acetic    acid ethyl ester;-   3-{4-Methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidin-1-yl}-3-oxo-propionitrile;-   3,3,3-Trifluoro-1-{4-methyl-3-[methyl-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidin-1-yl}-propan-1-one;-   1-{4-Methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidin-1-yl}-but-3-yn-1-one;-   1-{3-[(5-Chloro-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-methyl-amino]-4-methyl-piperidin-1-yl}-propan-1-one;-   1-{3-[(5-Fluoro-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-methyl-amino]-4-methyl-piperidin-1-yl}-propan-1-one;-   N-cyano-4-methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-N′-propyl-piperidine-1-carboxamidine;    and-   N-cyano-4,N′,N′-Trimethyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidine-1-carboxamidine.

In a particular embodiment, the Jak3 inhibitor is3-{(3R,4R)-4-Methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidin-1-yl}-3-oxo-propionitrile(CP0690550), which has the structure:

In one aspect of the invention, administration of the Jak3 inhibitorsignificantly increases tear production volume as compared to untreatedtear production volume within two days. In a further aspect of theinvention, administration of the Jak3 inhibitor increases tearproduction volume by at least 30% over initial tear production withinthe first two days of administration. In a further aspect,administration of the Jak3 inhibitor increases tear production volume byat least 50% over initial tear production within the first two days ofadministration. In a further aspect of the invention, administration ofthe Jak3 inhibitor increases tear production volume by at least 100%over initial tear volume within the first two days of administration. Inanother embodiment, the Jak3 inhibitor increases tear production volumeto a level comparable to that of normal tear production. In a furtheraspect of this embodiment, the Jak3 inhibitor increases tear productionvolume to a level comparable to that of normal tear production within 8days of initial treatment.

In an alternative embodiment, the invention comprises restoration ofnormal tear production in a mammal comprising administering to a mammala composition comprising a pharmaceutically acceptable carrier and apharmaceutically effective amount of a Janus kinase-3 (“Jak3”)inhibitor. In one aspect of this embodiment, the JAK3 inhibitor is3-{(3R,4R)-4-Methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidin-1-yl}-3-oxo-propionitrile(CP0690550).

In another aspect of the invention, the Jak3 inhibitor is administeredat most twice a day (BID). In a further aspect, the Jak 3 inhibitor isadministered daily (QD).

In a further aspect of the invention, the composition is topicallyadministered to the eye.

In another aspect of the invention, the pharmaceutically effectiveamount of the Jak3 inhibitor is 0.0001% to less than 1.0% (w/w). In afurther aspect the amount of the Jak3 inhibitor is 0.0003% to less than0.1% (w/w). In another aspect of the invention, the amount of the Jak3inhibitor is 0.0003 to 0.03% (w/w). In still another aspect the amountof the Jak3 inhibitor is 0.003% to 0.005% (w/w). In still anotheraspect, the amount of the Jak3 inhibitor is 0.01% to 0.03% (w/w). In astill further embodiment, the amount of Jak3 inhibitor is about 0.003%,0.005%, 0.01% or 0.03% (w/w).

In another aspect of the invention, the topical ophthalmic compositionfurther comprises a tonicity agent and a buffer. In a still anotheraspect of the invention, the tonicity agent is either a simple sugar ora sugar alcohol. In still another aspect of the invention, the buffer isselected from phosphate or citrate.

In still another aspect of the invention, the composition furthercomprises a surfactant. In a preferred embodiment of the invention, thesurfactant is selected from TritonX114 and tyloxapol. In further aspectof this embodiment, the composition further comprises a stabilizingpolymer. In a still further aspect of the invention, the stabilizingpolymer is carbomer 974p.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows tear production of the MRL/Ipr mouse (“dry eye mouse”)versus the control C57 mouse as discussed in Example 1 below.

FIG. 2 shows tear production of the dry eye mouse versus the C57 mouse,as when administered with the selective Jak3 inhibitor,3-{(3R,4R)-4-Methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidin-1-yl}-3-oxo-propionitrile(CP0690550).

FIG. 3 shows tear production of the Jak3 inhibitor CP0690550 as comparedto the leading dry eye therapeutic Restasis®.

FIG. 4 shows the dose response effects of CP690550 at 0.003%, 0.01%, and0.03% (w/w) in Formulation 1 vehicle as compared to normal and dry eyecontrols.

FIG. 5 shows the dose response effects of CP690550 at 0.001% and 0.003%(w/w) in Formulation 3 vehicle, as compared to normal and dry eyecontrols.

FIG. 6 shows the dose response effects of CP690550 at 0.001%, 0.003% and0.01% (w/w) in Formulation 4 vehicle, as compared to normal and dry eyecontrols.

FIG. 7 shows the dose response effects of CP690550 at 0.003% and 0.01%(w/w) in Formulation 3 vehicle in an induced dry eye mouse model, ascompared to normal eye and placebo vehicle controls.

FIG. 8 shows the dose response effects of CP690550 at 0.003% and 0.005%(w/w) in Formulation 5 vehicle in an induced dry eye mouse model, ascompared to normal eye and placebo vehicle controls.

DETAILED DESCRIPTION OF THE INVENTION

According to the present invention, inhibitors of Janus kinase-3 (Jak3),are administered to a mammal suffering from dry eye, and in particular,to a human patient suffering from dry eye. The Jak3 selective inhibitorsof the present invention in one aspect have selectivity for inhibitionof Jak3 relative to other protein tyrosine kinases, particularly theclosely related family member, Jak2. This is because Jak2 controlssignaling through the erythropoietin (EPO), macrophagecolony-stimulating factor (M-CSF), granulocyte-macrophage (GM)_CSF, andthrombopoietin (TPO) receptors. Moreover Jak2 deficiency isembryonically lethal due to impaired erythropoiesis. Significantpharmacologic inhibition of Jak2 in vivo could be expected to result inanemia, thrombocytopenia, and leukopenia. Pesu et al, ImmunologicalReviews 203, 127-142 (2005). At the enzymatic level, the Jak3 inhibitorsof the present invention are at least 10 fold more potent against Jak3versus Jak2, and are greater than 3000-fold specific relative to otherkinases.

The present invention relates to a method of treating dry eye comprisingadministering to a mammal a composition comprising a pharmaceuticallyacceptable carrier and a pharmaceutically effective amount of a Januskinase-3 (“Jak3”) inhibitor having the formula I and/or the specificembodiments as described above. Jak3 inhibitors are disclosed in U.S.Pat. Nos. 6,627,754 and 7,091,208, the contents of which areincorporated by reference herein. A particular embodiment of theinvention comprises administering to a patient a pharmaceuticallyeffective amount of3-{(3R,4R)-4-Methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidin-1-yl}-3-oxo-propionitrile.

The term “alkyl”, as used herein, unless otherwise indicated, includessaturated monovalent hydrocarbon radicals having straight or branchedmoieties or combinations thereof.

The term “alkoxy”, as used herein, includes O-alkyl groups wherein“alkyl” is defined above.

The term “halo”, as used herein, unless otherwise indicated, includesfluoro, chloro, bromo or iodo.

The compounds of this invention may contain double bonds. When suchbonds are present, the compounds of the invention exist as cis and transconfigurations and as mixtures thereof.

Unless otherwise indicated, the alkyl and alkenyl groups referred toherein, as well as the alkyl moieties of other groups referred to herein(e.g., alkoxy), may be linear or branched, and they may also be cyclic(e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl)or be linear or branched and contain cyclic moieties. Unless otherwiseindicated, halogen includes fluorine, chlorine, bromine, and iodine.

(C₂-C₉)Heterocycloalkyl when used herein refers to pyrrolidinyl,tetrahydrofuranyl, dihydrofuranyl, tetrahydropyranyl, pyranyl,thiopyranyl, aziridinyl, oxiranyl, methylenedioxyl, chromenyl,isoxazolidinyl, 1,3-oxazolidin-3-yl, isothiazolidinyl,1,3-thiazolidin-3-yl, 1,2-pyrazolidin-2-yl, 1,3-pyrazolidin-1-yl,piperidinyl, thiomorpholinyl, 1,2-tetrahydrothiazin-2-yl,1,3-tetrahydrothiazin-3-yl, tetrahydrothiadiazinyl, morpholinyl,1,2-tetrahydrodiazin-2-yl, 1,3-tetrahydrodiazin-1-yl,tetrahydroazepinyl, piperazinyl, chromanyl, etc. One of ordinary skillin the art will understand that the connection of said(C₂-C₉)heterocycloalkyl rings is through a carbon or a sp³ hybridizednitrogen heteroatom.

(C₂-C₉)Heteroaryl when used herein refers to furyl, thienyl, thiazolyl,pyrazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyrrolyl, triazolyl,tetrazolyl, imidazolyl, 1,3,5-oxadiazolyl, 1,2,4-oxadiazolyl,1,2,3-oxadiazolyl, 1,3,5-thiadiazolyl, 1,2,3-thiadiazolyl,1,2,4-thiadiazolyl, pyridyl, pyrimidyl, pyrazinyl, pyridazinyl,1,2,4-triazinyl, 1,2,3-triazinyl, 1,3,5-triazinyl,pyrazolo[3,4-b]pyridinyl, cinnolinyl, pteridinyl, purinyl,6,7-dihydro-5H-[1]pyrindinyl, benzo[b]thiophenyl,5,6,7,8-tetrahydro-quinolin-3-yl, benzoxazolyl, benzothiazolyl,benzisothiazolyl, benzisoxazolyl, benzimidazolyl, thianaphthenyl,isothianaphthenyl, benzofuranyl, isobenzofuranyl, isoindolyl, indolyl,indolizinyl, indazolyl, isoquinolyl, quinolyl, phthalazinyl,quinoxalinyl, quinazolinyl, benzoxazinyl; etc. One of ordinary skill inthe art will understand that the connection of said(C₂-C₉)heterocycloalkyl rings is through a carbon atom or a sp³hybridized nitrogen heteroatom.

(C₆-C₁₀)aryl when used herein refers to phenyl or naphthyl.

Preferred compounds of formula I include those wherein a is 0; b is 1; Xis carbonyl; c is 0; d is 0; e is 0; f is 0; and g is 0. Additionalpreferred compounds of formula I include those wherein a is 0; b is 1; Xis carbonyl; c is 0; d is 1; e is 0; f is 0, and g is 0. Additionalpreferred compounds of formula I include those wherein a is 0; b is 1; Xis carbonyl; c is 1; d is 0; e is 0; f is 0; and g is 0. Additionalpreferred compounds of formula I include those wherein a is 0; b is 1; Xis —C(═N=cyano)-; c is 1; d is 0; e is 0; f is 0; and g is 0.

Additional preferred compounds of formula I include those wherein a is0; b is 0; c is 0; d is 0; e is 0; f is 0; g is 1; and Z is —C(O)—O—.Additional preferred compounds of formula I include those wherein a is0; b is 1; X is S(O)_(n); n is 2; c is 0; d is 0; e is 0; f is 0; and gis 0. Additional preferred compounds of formula I include those whereina is 0; b is 1; X is S(O)_(n); n is 2; c is 0; d is 2; e is 0; f is 1; gis 1; and Z is carbonyl. Additional preferred compounds of formula Iinclude those wherein a is 0; b is 1; X is S(O)_(n); n is 2; c is 0; dis 2; e is 0; f is 1; and g is 0. Additional preferred compounds offormula I include those wherein a is 0; b is 1; X is carbonyl; c is 1; dis 0; e is 1; Y is S(O)_(n); n is 2; f is 0; and g is 0. Additionalpreferred compounds of formula I include those wherein a is 0; b is 1; Xis S(O)_(n); n is 2; c is 1; d is 0; e is 0; f is 0; and g is 0.Additional preferred compounds of formula I include those wherein a is1; b is 1; X is carbonyl; c is 1; d is 0; e is 0; f is 0; and g is 0.Additional preferred compounds of formula I include those wherein a is0; b is 1; X is S(O)_(n); c is 0; d is 1; e is 1; Y is S(O)_(n); n is 2;f is 0; and g is 0. Additional preferred compounds of formula I includethose wherein a is 0; b is 1; X is S(O)_(n); c is 0; d is 1; e is 1; Yis S(O)_(n); n is 2; f is 1; and g is 0. Additional preferred compoundsof formula I include those wherein a is 0; b is 1; X is oxygen; c is 0;d is 1; e is 1; Y is S(O)_(n); n is 2; f is 1; and g is 0. Additionalpreferred compounds of formula I include those wherein a is 0; b is 1; Xis oxygen; c is 0; d is 1; e is 1; Y is S(O)_(n); n is 2; f is 0; and gis 0. Additional preferred compounds of formula I include those whereina is 0; b is 1; X is carbonyl; c is 1; d is 1; e is 1; Y is S(O)_(n); fis 0; and g is 0. Additional preferred compounds of formula I includethose wherein a is 0; b is 1; X is carbonyl; c is 1; d is 1; e is 1; Yis S(O)_(n); n is 2; f is 1; and g is 0. Additional preferred compoundsof formula I include those wherein R¹² is cyano, trifluoromethyl,(C₁-C₆)alkyl, trifluoromethyl(C₁-C₆)alkyl, (C₁-C₆)alkylamino,((C₁-C₆)alkyl)₂amino, (C₂-C₆)alkynyl, cyano(C₁-C₆)alkyl,(C₁-C₆)alkyl-S(O)_(m) wherein m is 0, 1 or 2.

The compositions administered according to the present inventioncomprise a pharmaceutically effective amount of one or more of thespecified Jak3 inhibitors. As used herein, a “pharmaceutically effectiveamount” is one which is sufficient to reduce or eliminate signs ofsymptoms of dry eye or other disorders requiring the wetting of the eye.An effective dosage can be administered in one or more administrations.

As is understood in the clinical context, administration of an effectiveamount of a drug, compound, or pharmaceutical composition may or may notbe achieved in conjunction with another drug, compound, orpharmaceutical composition. Thus, an “effective amount” or “effectivedosage” may be considered in the context of administering one or moretherapeutic agents, and a single agent may be considered to be given inan effective amount if, in conjunction with one or more other agents, adesirable result may be or is achieved.

As used herein, “treatment” or “treating” is an approach for obtainingbeneficial or desired results including clinical results. For purposesof this invention, beneficial or desired clinical results include, butare not limited to, reducing and/or eliminating signs of symptoms of dryeye or other disorders requiring the wetting of the eye.

Most clinicians diagnose and treat Dry Eye Syndrome based on thesymptoms alone (JAMA 2001; 286:2114-9). Responses to the McMonnies & HoDry Eye Questionnaire can be used as a tool to assess the symptoms ofdry eye (score of more than 14.5 is consistent with dry eye diagnosis).Nonetheless, the National Eye Institute workshop on dry eyes defines‘symptoms of ocular discomfort’ as only one aspect of ‘dry eyes’. Ananalysis of demonstrable tear deficiency or possibility of excessivetear evaporation and damage to the exposed surface of the eye, canprovide confirmation of a ‘dry eye’ diagnosis.

As used herein, “significantly increases tear production” means astatistically significant (i.e. p<0.05) increase in tear production asmeasured by standard ophthalmic practice, i.e. Schirmer's or phenol redthreat test, fluorescein breakup time, or any of the tests describedbelow.

The most common tests for dry eye are tear film break-up time, Schirmertest and fluorescein staining, although these tests are non-exhaustive.The following are known tests for dry eye and the cutoff value at whichthey are considered abnormal and indicative of dry eyes:

TABLE 1 TEST Abnormal cutoff value for dry eye diagnosis Schirmer's ILess than or equal to 5 mm wetting over 5 min Phenol Red Thread Lessthan 9 mm Tear Breakup time Less than or equal to 10 seconds Fluoresceinstaining More than 3 out of 15 Rose Bengal staining More than 3 out of18 Tear film osmolarity Less than or equal to 312 mOsm/L Impressioncytology More than 1 Brush cytology More than 1 Tear lactoferrin Lessthan or equal to 0.9 ug/mL

Schirmer's I Test

In the Schirmer I test, the quantity of tears that are produced by theeye are measured: the tears are collected for approximately 5 minutes orso, to allow the ophthalmologist to determine whether the amountproduced is sufficient for maintaining eye health or not. If not muchtears are produced, then a tear deficient dry eye can be diagnosed. Ifenough tears are produced, but the patient still has symptoms of oculardiscomfort, then evaporative dry eye may be diagnosed.

In the Schirmer I test, a 35 mm×5 mm size filter paper strip is used tomeasure the amount of tears that are produced over a period of 5minutes. The strip is placed at the junction of the middle and lateralthirds of the lower eye lid under ambient light. The patient isinstructed to look forward and to blink normally during the course ofthe test. A negative test (more than 10 mm wetting of the filter paperin 5 minutes) means that the patient produces a normal quantity oftears. Patients with dry eyes have wetting values of less than 5 mm in 5minutes.

An important limitation of Schirmer test is that there may beconsiderable variability in the results of tests done at different timesand by different doctors. Its main utility may really be in diagnosingpatients with severe dry eyes.

There is some debate about the Schirmer I test. When an anestheticeyedrop is NOT used then this test is thought to measure basal andreflex tear secretion. When an anesthetic eyedrop IS used then this testis thought to measure only the basic tear secretion. There is compellingreason to believe that the tears measured by these two different methodsmay not sufficiently differentiate between basic and reflex tearproduction. Most clinicians perform this test after using anestheticeyedrops to numb the eye. However, The National Eye Institute workshopon dry eyes recommended not to use anesthetic eyedrops before performingthis test. The cutoff value is similar whether or not anesthetic isused. To measure the reflex tear secretion Schirmer II test may beperformed. Schirmer II test is performed by irritating the nasal mucosawith a cotton-tipped applicator prior to measuring tear production.

As an alternative to the Schirmer I or II test, which some cliniciansconsider unduly invasive and of little value for mild to moderate dryeyes, the phenol red thread test (Quick Zone®) can be used. A cottonthread impregnated with phenol red dye is used. Phenol red is pHsensitive and changes from yellow to red when wetted by tears. Thecrimped end of a 70 mm long thread is placed in the lower conjunctivalformix. After 15 seconds, the length of the color change on thethread—indicating the length of the thread wetted by the tears—ismeasured in millimeters. Wetting lengths should normally be between 9 mmand 20 mm. Patients with dry eyes have wetting values of less than 9 mm.

Tear Breakup Time (BUT)

In patients with dry eyes the tear film is unstable, and breaks upfaster. Therefore the tear break up time in patients who have dry eyesis shorter. Fluorescein BreakUp Time (FBUT) is used most commonly. Astrip of fluorescein is applied in the lower eyelid formix and thenremoved. The patient is asked to blink three times and then lookstraight forward, without blinking. The tear film is observed undercobalt-blue filtered light of the slitlamp microscope and the time thatelapsed between the last blink and appearance of the first break in thetear film is recorded with a stopwatch (a break is seen as a dark spotin a sea of blue). Fluorescein BUT of less than 10 seconds or less isconsistent with dry eyes.

To overcome limitations caused by the potential invasiveness of the FBUTtest, Non Invasive Break up time (NIBUT) methods have been developed.They are called Non Invasive because the eye is not touched. Instrumentssuch as a keratometer, hand-held keratoscope or Tearscope are requiredto measure NIBUT. A prerupture phase that precedes actual break up ofthe tear film can also be observed with some techniques. Thispre-rupture phase is termed Tear Thinning Time (TTT). Measurement isachieved by observing the distortion (TTT) and/or break up (NIBUT) of akeratometer mire (the reflected image of keratometer grid). Theclinician focuses and views the crisp mires, and then records the timetaken for the mire image to distort (TTT) and/or break up (NIBUT). NIBUTmeasurements are longer than fluorescein break up time. NIBUT values ofless than 15 seconds are consistent with dry eyes. TTT/NIBUT areconsidered to be more patient-friendly, repeatable and precise.

As used herein, administration “in conjunction” includes simultaneousadministration and/or administration at different times. Administrationin conjunction also encompasses administration as a co-formulation oradministration as separate compositions. As used herein, administrationin conjunction is meant to encompass any circumstance wherein a Jak3inhibitor and another agent are administered to an indivisual, which canoccur simultaneously and/or separately. As further discussed herein, itis understood that a Jak3 inhibitor and the other agent can beadministered at different dosing frequencies or intervals. For example,the Jak3 inhibitor can be administered daily, while the other agent canbe administered less frequently. It is understood that the Jak3inhibitor and the other agent can be administered using the same routeof administration or different routes of administration.

An “individual” (alternatively referred to as a “subject”) is a mammal,more preferably a human. A “mammal” includes, but is not limited to,farm animals (such as cows), sport animals, pets (such as cats, dogs,horses), primates, mice and rats.

The term “restoration of normal tear production” refers to the cessationof dry eye symptoms as described in standard ophthalmic practice, suchas a response score of less than 14.5 in McMonnies & Ho Dry EyeQuestionnaire and/or test results (i.e. from Schirmer's, red phenol,fluorescein etc as described in Table 1) which are in the normalvariable range.

According to the methods of the present invention, a compositioncomprising one or more of the specified Jak3 inhibitors and apharmaceutically acceptable carrier for topical ophthalmicadministration or implantation into the conjunctival sac or anteriorchamber of the eye is administered to a mammal in need thereof. Thecompositions are formulated in accordance with methods known in the artfor the particular route of administration desired.

Generally, for compositions intended to be administered topically to theeye in the form of eye drops or eye ointments, the total amount of theJak3 inhibitor will be about 0.0001 to less than 1.0% (w/w). A preferredrange of the amount of the Jak3 inhibitor is 0.0003% to less than 0.1%(w/w), while an even more preferred range of the amount of the Jak3inhibitor is 0.003 to 0.03% (w/w). Another preferred range of the amountof Jak3 inhibitor is 0.005% to 0.05% (w/w), and a still more preferredrange is 0.01% to 0.03% (w/w).

Preferably, the compositions administered according to the presentinvention will be formulated as solutions, suspensions, emulsions andother dosage forms for topical administration. Aqueous solutions aregenerally preferred, based on ease of formulation, as well as apatient's ability to easily administer such compositions by means ofinstilling one to two drops of the solutions in the affected eyes.However, the compositions may also be suspensions, viscous orsemi-viscous gels, or other types of solid or semi-solid compositions.Suspensions may be preferred for Jak3 inhibitors which are sparinglysoluble in water.

The compositions administered according to the present invention mayalso include various other ingredients, including but not limited totonicity agents, buffers, surfactants, stabilizing polymer,preservatives, co-solvents and viscosity building agents. Preferredpharmaceutical compositions of the present invention include theinhibitor with a tonicity agent and a buffer. The pharmaceuticalcompositions of the present invention may further optionally include asurfactant and/or a palliative agent and/or a stabilizing polymer.

Various tonicity agents may be employed to adjust the tonicity of thecomposition, preferably to that of natural tears for ophthalmiccompositions. For example, sodium chloride, potassium chloride,magnesium chloride, calcium chloride, simple sugars such as dextrose,fructose, galactose, and/or simply polyols such as the sugar alcoholsmannitol, sorbitol, xylitol, lactitol, isomalt, maltitol, andhydrogenated starch hydrolysates may be added to the composition toapproximate physiological tonicity. Such an amount of tonicity agentwill vary, depending on the particular agent to be added. In general,however, the compositions will have a tonicity agent in an amountsufficient to cause the final composition to have an ophthalmicallyacceptable osmolality (generally about 150-450 mOsm, preferably 250-350mOsm and most preferably at approximately 290 mOsm). In general thetonicity agents of the invention will be present in the range of 2 to 4%w/w. Preferred tonicity agents of the invention include the simplesugars or the sugar alcohols. A particular embodiment of the inventionis D-mannitol.

An appropriate buffer system (e.g., sodium phosphate, sodium acetate,sodium citrate, sodium borate or boric acid) may be added to thecompositions to prevent pH drift under storage conditions. Theparticular concentration will vary, depending on the agent employed.Preferably however, the buffer will be chosen to maintain a target pHwithin the range of pH 5 to 8, and more preferably to a target pH of pH5 to 7.

Surfactants may optionally be employed to deliver higher concentrationsof inhibitor. The surfactants function to solubilize the inhibitor andstabilize colloid dispersion, such as micellar solution, microemulsion,emulsion and suspension. Examples of surfactants which may optionally beused include polysorbate, poloxamer, polyosyl 40 stearate, polyoxylcastor oil, tyloxapol, triton, and sorbitan monolaurate. Preferredsurfactants to be employed in the invention have ahydrophile/lipophile/balance “HLB” in the range of 12.4 to 13.2 and areacceptable for ophthalmic use, such as TritonX114 and tyloxapol.

Compositions formulated for the treatment of dry eye-type diseases anddisorders may also comprise aqueous carriers designed to provideimmediate, short-term relief of dry eye-type conditions. Such carrierscan be formulated as a phospholipid carrier or an artificial tearscarrier, or mixtures of both. As used herein, “phospholipid carrier” and“artificial tears carrier” refer to aqueous compositions which: (i)comprise one or more phospholipids (in the case of phospholipidcarriers) or other compounds, which lubricate, “wet”, approximate theconsistency of endogenous tears, aid in natural tear build-up, orotherwise provide temporary relief of dry eye symptoms and conditionsupon ocular administration; (ii) are safe; and (iii) provide theappropriate delivery vehicle for the topical administration of aneffective amount of one or more of the specific Jak3 inhibitors.Examples or artificial tears compositions useful as artificial tearscarriers include, but are not limited to, commercial products, such asVisine Pure Tears®, Visine Tears Natural Tears Formula® (Johnson &Johnson) Tears Naturale®, Tears Naturale II®, Tears Naturale Free®, BionTears® (Alcon Laboratories, Inc. Forth Worth, Tex.), Refresh Tears®,Refresh Endura®, Refresh Plus® (Allergan Inc.). Examples of phospholipidcarrier formulations include those disclosed in U.S. Pat. Nos. 4,804,539(Guo et al.), 4,883,658 (Holly), 4,914,088 (glonek), 5,075,104 (Gresselet al.), 5,278,151 (Korb et al.), 5,294,607 (Glonek et al.), 5,371,108(Korb et al.), 5,578,586 (Glonek et al.); the foregoing patents areincorporated herein by reference to the extent they disclosephospholipid compositions useful as phospholipid carriers of the presentinvention.

Palliative compounds designed to lubricate, “wet,” approximate theconsistency of endogenous tears, aid in natural tear build-up, orotherwise provide temporary relief of dry eye symptoms and conditionsupon ocular administration of the eye are known in the art. Suchpalliative agents may, in some cases, serve a dual function as atonicity agent, such as the monomeric polyols (glycerol, propyleneglycol, ethylene glycol). The palliative compounds may enhance theviscosity of the composition, and include, but are not limited to:monomeric polyols, polymeric polyols, such as polyethylene glycol,hydroxypropylmethyl cellulose (“HPMC”), carboxy methylcellulose sodium,hydroxyl propylcellulose (“HPC”), dextrans, such as dextran 70; watersoluble proteins, such as gelatin; and vinyl polymers, such as polyvinylalcohol, polyvinylpyrrolidinone, povidone, and carbomers, such as,carbomer 941, carbomer 940, carbomer 971P, carbomer 974P.

Additional agents that may be added to the ophthalmic compositions ofthe present invention are demulcents which function as a stabilizingpolymer. The stabilizing polymer should be an ionic/charged example withprecedence for topical ocular use, more specifically a polymer thatcarries negative charge on it's surface that can exhibit azeta-potential of (−)10-50 mV for physical stability and capable ofmaking a dispersion in water (i.e. water soluble). A preferredstabilizing polymer of the invention would be polyelectrolyte orpolyectrolytes if more than one, from the family of cross-linkedpolyacrylates, such as carbomers and Pemulen®, specifically Carbomer974p (polyacrylic acid), at 0.1-0.5% w/w. As noted above, the carbomersmay also function as a palliative compound.

Other compounds may also be added to the ophthalmic compositions of thepresent invention to increase the viscosity of the carrier. Examples ofviscosity enhancing agents include, but are not limited to:polysaccharides, such as hyaluronic acid and its salts, chondroitinsulfate and its salts, dextrans, various polymers of the cellulosefamily; vinyl polymers; and acrylic acid polymers. In general, thephospholipid carrier or artificial tears carrier compositions willexhibit a viscosity of 1 to 400 centipoises (“cps”).

Topical ophthalmic products are typically packaged in multidose form.Preservative are thus required to prevent microbial contamination duringuse. Suitable preservatives include: benzalkonium chloride,chlorobutanol, benzododecinium bromide, methyl paraben, propyl paraben,phenylethyl alcohol, edentate disodium, sorbic acid, polyquaternium-1,or other agents known to those skilled in the art. Such preservativesare typically employed at a level of from 0.001 to 1.0% w/v. Unit dosecompositions of the present invention will be sterile, but typicallyunpreserved. Such compositions, therefore, generally will not containpreservatives.

The preferred compositions of the present invention are intended foradministration to a human patient suffering from dry eye or symptoms ofdry eye. Preferably, such compositions will be administered topically.In general, the doses used for the above described purposes will barybut will be in an effective amount to eliminate or improve dry eyeconditions. Generally, 1-2 drops of such compositions will beadministered from once to many times per day. Preferably suchcompositions will be administered once (QD) or twice (BID) daily.

Representative eye drop formulations are provided in Example 2 below.

Example 11-{4-Methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidin-1-yl}-ethanoneand3-{(3R,4R)-4-Methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidin-1-yl}-3-oxo-propionitrile.Method A: (1-Benzyl-4-methyl-piperidin-3-yl)-methyl-amine

To a stirred solution of 1-benzyl-4-methyl-piperidin-3-one (2.3 grams,11.5 mmol), prepared by the methods of Iorio, M. A. and Damia, G.,Tetrahedron, 26, 5519 (1970) and Grieco et al., Journal of the AmericanChemical Society, 107, 1768 (1985), (modified using 5% methanol as aco-solvent), both references are incorporated by reference in theirentirety, dissolved in 23 mL of 2 M methylamine in tetrahydrofuran wasadded 1.4 mL (23 mmol) of acetic acid and the resulting mixture stirredin a sealed tube for 16 hours at room temperature. Triacetoxy sodiumborohydride (4.9 grams, 23 mmol) was added and the new mixture stirredat room temperature in a sealed tube for 24 h, at which time, thereaction was quenched upon addition of 1 N sodium hydroxide (50 mL). Thereaction mixture was then extracted 3×80 mL with ether, the combinedether layers dried over sodium sulfate (Na₂SO₄) and concentrated todryness in vacuo affording 1.7 grams (69%) of the title compound as awhite solid. LRMS: 219.1 (M+1).

Method B:(1-Benzyl-4-methyl-piperidin-3-yl)-methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amine

A solution of 4-chloropyrrolo[2,3-d]pyrimidine (2.4 grams, 15.9 mmol),prepared by the method of Davoll, J. Am. Chem. Soc., 82, 131 (1960),which is incorporated by reference in its entirety, and the product fromMethod A (1.7 grams, 7.95 mmol) dissolved in 2 equivalents oftriethylamine was heated in a sealed tube at 100° C. for 3 days.Following cooling to room temperature and concentration under reducedpressure, the residue was purified by flash chromatography (silica; 3%methanol in dichloromethane) affording 1.3 grams (50%) of the titlecompound as a colorless oil. LRMS: 336.1 (M+1).

Method C:Methyl-(4-methyl-piperidin-3-yl)-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amine

To the product from Method B (0.7 grams, 2.19 mmol) dissolved in 15 mLof ethanol was added 1.5 mL of 2 N hydrochloric acid and the reactionmixture degassed by nitrogen purge. To the reaction mixture was thenadded 0.5 grams of 20% palladium hydroxide on carbon (50% water)(Aldrich) and the resulting mixture shaken (Parr-Shaker) under a 50 psiatmosphere of hydrogen at room temperature for 2 days. The Celitefiltered reaction mixture was concentrated to dryness in vacuo and theresidue purified by flash chromatography (silica; 5% methanol indichoromethane) affording 0.48 grams (90%) of the title compound. LRMS:246.1 (M+1).

Method D:1-{4-Methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidin-1-yl}-ethanone

To a stirred solution of the product from Method C (0.03 grams, 0.114mmol) dissolved in 5 mL of 10:1 dichloromethane/pyridine was added(0.018 grams, 0.228 mmol) of acetylchloride and the resulting mixturestirred at room temperature for 18 hours. The reaction mixture was thenpartitioned between dichloromethane and saturated sodium bicarbonate(NaHCO₃). The organic layer was washed again with saturated NaHCO₃,dried over sodium sulfate and concentrated to dryness in vacuo. Theresidue was purified by preparative thin layer chromatography (PTLC)(silica; 4% methanol in dichloromethane) affording 0.005 mg (15%) of thetitle compound as a colorless oil. LRMS: 288.1 (M+1).

Method E: Synthesis of3-{(3R,4R)-4-Methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidin-1-yl}-3-oxo-propionitrile(CP0690550)

The title compound3-{(3R,4R)-4-Methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino]-piperidin-1-yl}-3-oxo-propionitrile(CP0690550) was prepared using routine modification of the Methods A-Ddescribed above.

Method F: Preparation of3-{(3R,4R)-4-Methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidin-1-yl}-3-oxo-propionitrilecitrate salt

The citrate salt was prepared using methods described in WO2007/012953.To a clean, dry, nitrogen-purged 500 ml reactor were chargedmethyl-4(-methyl-piperidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amine(25.0 g, 0.102 mol) and methylene chloride (250 ml). The mixture wasstirred at room temperature for a minimum of 2.5 hours. To a clean, dry,nitrogen-purged 1 L reactor were charged cyanoacetic acid (18.2 g, 0.214mol), methylene chloride (375 ml), and triethyl amine (30.1 ml, 0.214mol). The mixture was cooled to −15-5.0° C. over one hour andtrimethylacetyl chloride (25.6 ml, 0.204 mol) was added at a rate tomaintain the temperature below 0° C. The reaction was held for a minimumof 2.5 hours, then the solution of the amine was added at a rate thatmaintained the temperature below 0° C. After stirring for 1 hour, themixture was warmed to room temperature and 1M sodium hydroxide (125 ml)was added. The organic layer was washed with water (125 ml). Themethylene chloride solution was displaced with acetone until a volume of500 ml and a temperature of 55-65° C. had been achieved. Water (75 ml)was charged to the mixture while maintaining the temperature at 55-65°C. A solution of citric acid (20.76 g, 0.107 mol) in water (25.0) wascharged and the mixture was cooled to room temperature. The reactor wasstirred for a minimum of 5 hours and then the resulting solids wereisolated by filtration and washed with acetone (2×75 ml), which was sentto the filter. The salt was charged into a clean, dry, nitrogen-purged 1L reactor with 2B ethanol (190 ml) and water (190 ml). The slurry washeated to 75-85° C. for a minimum of 4 hours. The mixture was cooled to20-30° C. and stirred for an additional 4 hours. The solids wereisolated by filtration and washed with 2B ethanol (190 ml). After dryingin a vacuum oven at 50° C. with a slight nitrogen bleed, 34.6 g (67.3%)of the title compound were isolated.

¹H NMR (500 MHz, d₆-DMSO): δ 8.14 (s, 1H, 7.11 (d, J=3.6 Hz, 1H), 6.57(d, J=3.6 Hz, 1H), 4.96 (q, J=6.0 Hz, 1H), 4.00-3.90 (m, 2H), 3.80 (m,2H), 3.51 (m, 1H), 3.32 (s, 3H), 2.80 (Ab_(q), J=15.6 Hz, 2H), 2.71(Abq, J=15.6 Hz, 2H), 2.52-2.50 (m, qH), 2.45-2.41 (m, 1H), 1.81 (m,1H), 1.69-1.65 (m, 1H), 1.04 (d, J=6.9 Hz, 3H).

Method G: Preparation of free base3-{(3R,4R)-4-Methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidin-1-yl}-3-oxo-propionitrile

The3-{(3R,4R)-4-Methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidin-1-yl}-3-oxo-propionitrilecitrate salt, as prepared in Method F above, was treated with 1M sodiumhydroxide in dichloromethane to afford the corresponding free base. Thefree base was then crystallized from methanol and water to furnish thecrystal form of the free base.

Example 2 Pharmaceutical Formulations Example 2a Formulations

Table 2 below illustrates various pharmaceutical formulations comprisingthe inhibitor CP0690550.

TABLE 2 Formulation code Formulation Description Formulation 1 50 mM pH7.4 phosphate buffer, 0.05% Tween 80, 0.5% NaCl Formulation 2 50 mM pH7.4 Phosphate buffer, 0.36% HPMC, 0.2% Glycerin, 1% PEG400, 0.35% NaClFormulation 3 5 mM pH 7.4 phosphate buffer, 0.36% HPMC, 0.2% Glyerin, 1%PEG400, 5% Cremophor ELP, 4.3% Mannitol Formulation 4 10 mM pH 5.8citrate buffer, 4.5% Mannitol Formulation 5 10 mM pH 5.8 citrate buffer,4.2% Mannitol, 0.36% HPMC, 0.2% Glycerin Formulation 6 0.3% tyloxapol(surfactant, nonionic), 0.5% Carbopol974P (polymeric stabilizer,anionic), 2.25% D-mannitol, pH 6.5 phosphate buffer at 50 mMconcentration, 230 mOsm/kg Formulation 7 0.3% tyloxapol (surfactant,nonionic), 0.1% Carbopol974P (polymeric stabilizer, anionic), 2.25%D-mannitol, pH 6.5 phosphate buffer at 50 mM concentration, 230 mOsm/kg

Example 2b Preparation

Each of the above formulations 1 through 5 were prepared comprising theJak3 inhibitor CP0690550 in three dosage amounts: 0.001%, 0.003% and0.01% w/w. Formulation 5 was also prepared with the Jak3 inhibitorCP9690550 in the amount of 0.005% w/w (see FIG. 8). Formulations 6 and 7were prepared comprising Jak3 inhibitor in the amount of 0.03% w/w.

The above formulations were prepared by adding the required amount ofthe tonicity agent (hypromellose, or mannitol where present) to a flask,heating to about to about 50° C. in half the final volume of buffer(phosphate or citrate buffer as indicated). After heating the batchquantities of inhibitor CP690550, the additional excipients (glycerin,polyethylene glycol 400, where indicated) were weighed and dissolved.The batch quantity of the Jak3 inhibitor free base as a stock solutionwas measured and added. Purified water was added to q.s. to 100%. Themixture was stirred for five minutes to homogenize and then filteredthrough a sterilizing filter membrane into a sterile recipient. Ifnecessary, pH was adjusted by admixture of 1.0 NaOH.

Formulations 6 and 7 represent ophthalmic compositions in which higheramounts of the inhibitor CP690550 are present (0.03% w/w). In additionto a tonicity agent and buffer, such higher concentrate formulationswill also include a surfactant and optionally a stabilizing polymer.Preferred surfactants of the invention include TritonX114 and tyloxapol.Preferred stabilizing polymers include the carbomer 974p.

Preparation of Formulations 6 and 7 were prepared by dispersing thecarbomer first in the surfactant containing buffer at 10× of their finalconcentration (e.g. 3% tyloxapol in 50 mM phosphate buffer at pH6.5 with2.5% D-manitol and 5% Carbomer 974p). The Jak3 inhibitor CP690550 wasdispersed in the preconcentrate also at 10× of its final concentration.The mixture was then homogenized, with final formulation being obtainedby 10× dilution of filtered preconcentrate in a matching buffer.

Example 3 Effect of selective Jak3 inhibitor on Tear Production MRL/Ipr(Dry Eye) Mice

To study the level of tear production over time, 10 MRL/Ipr mice and 5of the age-matched C57 Black/6 control mice at 8 weeks of age werepurchased from Jackson Laboratory (Bar Harbor, Me., USA). The mousestrain MRL/Ipr has a mutation which predisposes it to dry eye similar toSjogren's Syndrome.

Baseline tear volume was established by measuring of tear volume 4 timesover one week to establish that dry eye occurred in MRL/Ipr mice versusC57 mice. Tear production was measured with cotton thread (Quick Zone®)held with forceps and applied to the ocular surface at the lateralcanthus for 15 to 30 seconds. Wetting of the thread was measured inmillimeters. Tear production measurements of MRL/Ipr mice versusC57control mice are shown in FIG. 1.

Animals (5-10 mice/group) were treated with 2 μl of either vehicle(phosphate buffered saline, PBS, containing 0.05% Tween 80) or 0.1%(w/w) CP-690550 twice a day for two weeks. C57 mice also receivedvehicle. Tear volume was measured on days 2, 3, and 6. The effect ofCP-690550 on tear production in MRL/Ipr Mice is shown in FIG. 2.

As shown in FIG. 2, CP0690550 was able to significantly increase tearproduction volume within the first 2 days of application. Notably,CP690550 increased tear production volume by over 100% over initial tearproduction of the dry eye mice within the first 2 days of application.

Example 4 Comparative data comparing CP-690550 to Restasis®

Tear measurement was done using phenol red impregnated thread (QuickZone®) and baseline measurement was performed before each drugapplication. Drugs or vehicles were applied at 2 μl, two times a day.

Vehicle-treated normal C57black/6 and MRL/Ipr dry eye mice were used asnormal control and dry eye control, respectively. The vehicle forCP690550 was phosphate buffered saline, PBS, containing 0.05% Tween 80.

The experiment to compare efficacy of CP690550 and Restasis® was doneusing Restasis® (cyclosporine 0.05%) and CP-690550 at 0.01% appliedtwice a day for 30 days. Control animals included normal C57Black/6mice, and MRL/Ipr mice, treated with PBS vehicle and MRL/Ipr micetreated with Refresh Endura® (vehicle for Restasis) (5-10 mice/group).Comparative tear production of CP-690550 and Restasis® is shown in FIG.3.

As shown in FIG. 3, CP690550 had a much faster response rate as comparedto Restasis® and showed a significant improvement in the volume of tearproduction over Restasis®

Moreover, both FIGS. 3 and 4 indicate that administration of CP690550ultimately increased tear production volume to a level comparable tothat of normal tear production (within 5 to 10% of tear production by“normal” C57mice).

Example 5 Dose response Rates of CP690550 in MRL/Ipr Mice

As with Example 4 above, vehicle-treated normal C57black/6 and MRL/Iprdry eye mice (5-10 mice/group) were used as normal control and dry eyecontrol, respectively. The vehicles for CP690550 were Formulations 1, 3and 4 as indicated in Example 2 above. The Jak3 inhibitor CP690550 wasapplied at 0.003% (w/w), 01% (w/w) and 0.03% (w/w), as calculated forfree base concentration. Treatment for Formulation 1 was carried for twoweeks, at 41, twice a day. Treatment for Formulations 3 and 4 werecarried for 2 weeks, at 1 μl, once a day.

FIG. 4 illustrates that within 8 days, administration of CP690550 inFormulation 1 vehicle at 0.01% (w/w) twice a day increased tearproduction to a level comparable to that of normal tear production(within 5 to 10% of tear production by C57 normal mice). Similarly,FIGS. 5 and 6 illustrate dose response rates of CP690550 in Formulations3 and 4 vehicle. In all comparisons, significant improvement in tearproduction was shown at 2 days after treatment.

Example 6 Induced Dry Eye Mouse Model

An injection was prepared comprising 2.5 mg/ml scopolamine (sigma) ininjectable saline (1-1.5 ml per animal). In normal C57 mice (200-250) ulof scopolamine was injected four times every 2.5 hrs in alternating hindquarters. The mice were placed in special cages (with holes in front andback) and placed in a hood. Fans were placed in front of each cage, andturned on for 16 hours over night for five days. Measurements were takendaily. At the end of 5 days, all animals were considered dry-induced.

As with Examples 4 and 5 above, animals were treated with drug orvehicle at 1 μl, once a day for 2 weeks. Vehicle-treated C57 black/6 andthe dry-induced mice (5-10 mice/group) were treated with Formulations 3and 5 as prepared in Example 2 above. FIGS. 7 and 8 illustrate that asignificant increase in tear production was shown at 2 days oftreatment. After 4 days of treatment, formulations comprising 0.01%(w/w) of CP0690550 inhibitor in Formulation 3 and 0.003% of CP690550inhibitor in Formulation 5 approximated normal tear production values.

This invention has been described by reference to certain preferredembodiments; however, it should be understood that it may be embodied inother specific forms or variations thereof without departing from itsspecial or essential characteristics. The embodiments described aboveare therefore considered to be illustrative in all respects and notrestrictive, the scope of the invention being indicated by the appendedclaims rather than by the foregoing description.

What is claimed is:
 1. A topical ophthalmic composition comprising apharmaceutically effective amount of a Jak3 inhibitor or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier.
 2. The topical ophthalmic composition of claim 1,wherein the Jak3 inhibitor is3-{(3R,4R)-4-Methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidin-1-yl}-3-oxo-propionitrile(CP0690550), and has the structure:


3. The ophthalmic composition of claim 2, wherein the pharmaceuticallyeffective amount of Jak3 inhibitor is 0.0001 to less than 0.1% (w/w). 4.The ophthalmic composition of claim 1, wherein the pharmaceuticallyeffective amount of Jak3 inhibitor is 0.003 to 0.03% (w/w).
 5. Theophthalmic composition of claim 1, wherein the pharmaceuticallyeffective amount of Jak3 inhibitor is 0.01 to 0.03% (w/w).
 6. Theophthalmic composition of claim 2, wherein the buffer maintains a targetpH within the range of pH 5 to
 8. 7. The ophthalmic composition of claim2, further comprising a tonicity agent and a buffer selected fromphosphate or citrate.
 8. The ophthalmic composition according to claim7, further comprising a surfactant.
 9. The ophthalmic compositionaccording to claim 8, wherein the surfactant is tyloxapol.
 10. A topicalophthalmic composition comprising a pharmaceutically effective amount ofthe Jak3 inhibitor3-{(3R,4R)-4-Methyl-3-[methyl-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidin-1-yl}-3-oxo-propionitrile(CP0690550), or a pharmaceutically acceptable salt thereof:

and a pharmaceutically acceptable carrier.
 11. The ophthalmiccomposition of claim 10, wherein the pharmaceutically effective amountof Jak3 inhibitor is 0.0001 to less than 0.1% (w/w).
 12. The ophthalmiccomposition of claim 10, wherein the pharmaceutically effective amountof Jak3 inhibitor is about 0.03% (w/w).
 13. The ophthalmic compositionof claim 11, wherein the buffer maintains a target pH within the rangeof pH 5 to
 8. 14. The ophthalmic composition of claim 13, furthercomprising a tonicity agent and a buffer selected from phosphate orcitrate, and a surfactant.
 15. The ophthalmic composition according toclaim 14, wherein the surfactant is tyloxapol.